5'-Atp → Phenindione

HRID 569712Notes

Reaction equation

HRID 569712 reaction equation
P([O:13][CH2:14][C@H:15]1[O:19][C@@H:18](N2C3N=CN=C(N)C=3N=C2)[C@H:17](O)[C@@H:16]1[OH:31])(OP(OP(O)(O)=O)(O)=O)(=O)O.[CH3:32][C:33]1[C:43](=O)[C:42]2[CH:41]=[CH:40][CH:39]=[CH:38][C:37]=2[C:35](=[O:36])[C:34]=1[CH2:45]/[CH:46]=[C:47](/CCC[C@@H](CCC[C@@H](CCCC(C)C)C)C)\[CH3:48].[OH-:65].[Na+]>[Cl-].[Na+]>[CH:32]1[CH:40]=[CH:41][C:42]([CH:37]2[C:35](=[O:36])[C:34]3[CH:45]=[CH:46][CH:47]=[CH:48][C:39]=3[C:38]2=[O:65])=[CH:43][CH:33]=1.[CH3:46][C:45]([CH2:34][CH:35]([C:15]1[C:14](=[O:13])[O:19][C:18]2[CH:17]=[CH:16][CH:15]=[CH:14][C:17]=2[C:16]=1[OH:31])[C:37]1[CH:42]=[CH:41][CH:40]=[CH:39][CH:38]=1)=[O:65]

Participants

6

Reactant 3

HCID 59575'-AtpHCID 14798Sodium HydroxideHCID 5284607Phylloquinone

Product 2

HCID 4760PhenindioneHCID 54678486Warfarin

Reagents, catalysts & solvents 1

HCID 5234Sodium ChlorideSolvent

Procedure

Control, blank, and experimental reaction mixtures were made for each experiment. Control and blank reaction mixtures were made up with vitamin K and without vitamin K, respectively. Experimental reaction mixtures contained both vitamin K and a vitamin K antagonist. The control reaction mixture contained 3.00 ml of microsomal suspension, 0.81 ml of buffer II, 1.20 ml of an ATP-generating system (final concentrations: 1 mM ATP, 10 mM phosphocreatine, 2.5 mM Mg[acetate]2 ·4H2O, 20 μg/ml creatine phosphokinase [52 U/ml]), 0.60 ml of NADH (final concentration 2 mM), 0.30 ml of dithiothreitol (final concentration 7 mM) dissolved in buffer II, 0.060 ml of NaH14CO3 (1.0 μCi/μl; added 0.5 min prior to reaction initiation), and at the time of reaction initiation 0.030 ml of vitamin K1 (final concentration 20 μg/ml) diluted in 0.85% sodium chloride solution. This vitamin K concentration is associated with maximal in vitro incorporation of added H14CO3 into vitamin K-dependent substrate proteins (J. Biol. Chem. 251, 2770-2776). The blank reaction mixture consisted of the same components except that vitamin K was replaced by an equal volume of isotonic saline. The experimental reaction mixtures were made up in the same way as the control reaction mixtures with the exceptions that: (1) a volume of buffer II was replaced by a volume of buffer II containing an inhibitor of vitamin K-dependent carboxylation, and (2) all individual volumes were two-thirds of those in the control reaction mixtures. The sodium salts of TCP, phenindione, and warfarin were formed in aqueous sodium hydroxide solution (J. Am. Chem. Soc. 83, 2676-2679) and were freely soluble in buffer II at all concentrations used in these studies. Chloro-K1 was formulated as an oil-in-water emulsion with Tween 80 (5% v/v final concentration), while chloro-K3 was formulated as a suspension in Tween 80 (5% v/v final concentration) (Molecular Pharmacology 10, 373-380).

Workup

  1. customControl, blank, and experimental reaction mixtures
  2. customControl and blank reaction mixtures
  3. customExperimental reaction
  4. customThe control reaction mixture
  5. concentrationconcentrations
  6. dissolution1 mM ATP, 10 mM phosphocreatine, 2.5 mM Mg[acetate]2 ·4H2O, 20 μg/ml creatine phosphokinase [52 U/ml]), 0.60 ml of NADH (final concentration 2 mM), 0.30 ml of dithiothreitol (final concentration 7 mM) dissolved in buffer II, 0.060 ml of NaH14CO3 (1.0 μCi/μl
  7. additionadded 0.5 min
  8. customto reaction initiation), and
  9. concentrationThis vitamin K concentration
  10. additionof added H14CO3 into vitamin K-dependent substrate proteins (J. Biol. Chem. 251, 2770-2776)
  11. customThe blank reaction mixture
  12. customThe experimental reaction mixtures
  13. customwere made up in the same way as the control reaction mixtures with the exceptions that
  14. customin the control reaction mixtures

Sources & evidence

ORD record
ord-93c8b0ee552449dd89b1c1d008278304
Source dataset
uspto-grants-1988_03
Patent number
US04734282

Reaction notes

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